检出限
亚甲蓝
铁氰化物
电化学
DNA
化学
亚铁氰化物
小RNA
微乳液
三元运算
组合化学
生物物理学
电极
基因
生物化学
生物
色谱法
计算机科学
光催化
物理化学
程序设计语言
催化作用
肺表面活性物质
作者
Liangliang Wang,Xiufeng Wu,Guanyu Chen,Yawen Chen,Lilan Xu,Jingdong Wang,Jinghua Chen
标识
DOI:10.1016/j.bios.2023.115783
摘要
Exosomal microRNA (miRNA) are important biomarkers for liquid biopsy, and display clinical molecular signatures for cancer diagnosis. Although advanced detection methods have been established to detect exosomal miRNAs, they are faced with certain challenges. Therefore, we aimed to establish a dual amplification-based electrochemical method for detecting exosomal miRNA. This method combined a two-hairpins-based ternary hybridization structure (thTHS)-initiated single-stranded DNA (ssDNA) amplification reaction (ssDAR) and sodium perchlorate (NaClO4)-assisted electrocatalytic cycle. Two DNA hairpins were designed to hybridize with target miRNA, forming thTHS. Next, ssDAR was triggered by thTHS to produce long ssDNA on magnetic beads. The long ssDNA, complementary to the signal probes, was subsequently released onto a methylene blue (MB)-labeled double-stranded DNA-modified electrode for strand displacement reaction. This led to a quantitative change in MB and a change in electrocatalytic reduction current from the electrocatalytic cycle of MB-ferricyanide. An amplified electrocatalytic reduction current was produced by adding NaClO4 to the electrocatalytic system, which substantially improved the signal response range and detection sensitivity. Ultimately, exosomal miRNA detection was achieved by recording changes in the electrocatalytic reduction current before and after miRNA addition. This electrochemical method exhibited a sensitive concentration response with a detection limit of 45 aM and selective miRNA recognition, and successfully used to detect exosomal miRNA derived from cells and serum. Additionally, this method exhibited better discrimination ability between patients with breast cancer (BC) and those people without BC (patients with benign breast disease and healthy people), providing a promising strategy for detecting and monitoring cancer biomarkers.
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